Cloning and correct expression in Escherichia coli of the petE and petJ genes respectively encoding plastocyanin and cytochrome c6 from the cyanobacterium Anabaena sp. PCC 7119.
The genes coding for plastocyanin (petE) and cytochrome c6 (petJ) from Anabaena sp. PCC 7119 have been cloned and properly expressed in Escherichia coli. The recombinant proteins are identical to those purified from the cyanobacterial cells. The products of both the petE and petJ genes are correctly processed in E. coli, as deduced from their identical N-terminal amino acid sequences as compared with those of the metalloproteins isolated from the cyanobacterium. Physicochemical and functional properties of the native and recombinant protein preparations are also identical, thereby confirming that expression of petE and petJ genes in E. coli is an adequate tool to address the study of the structure/function relationships in plastocyanin and cytochrome c6 from Anabaena by site-directed mutagenesis.
Molina-Heredia
F P
FP
Instituto de Bioquímica Vegetal y Fotosíntesis, Centro Isla de la Cartuja, Universidad de Sevilla, Spain.
Hervás
M
M
Navarro
J A
JA
De la Rosa
M A
MA
D000595:Amino Acid Sequence; D017033:Anabaena / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003574:Cytochrome c Group / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003580:Cytochromes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D045348:Cytochromes f; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010970:Plastocyanin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013053:Spectrophotometry
D003574:Cytochrome c Group; D003580:Cytochromes; D017931:DNA Primers; D011994:Recombinant Proteins; C083951:cytochrome c553; D004247:DNA; D010970:Plastocyanin; D045348:Cytochromes f; C022302:cytochrome C-552